The goal of the current proposal is to apply state of the art gene modification technology including lentiviral delivery systems and RNA interference technology to commercially available tissue engineered human in vitro models. These models will consist of differentiated 3-D epidermal cultures and tracheal/bronchial epithelial cultures, as well as epidermal and airway epithelia co-cultured with mesenchymal cells. In Phase I research, the technology will be used to produce 3 types of genetically modified tissue engineered culture products: 1) stable addition of a functional gene, 2) stable silencing of a gene, and 3) stable introduction of a reporter for detection of gene activation. During Phase II, the techniques will be applied to additional tissue engineered human in vitro models, and the inventory of available gene modified products will be expanded. These proposed commercially available in vitro model systems will provide researchers in the pharmaceutical industry and academic research laboratories a readily available means for studying the functional genomics of nearly any gene in human epithelial cells, as well as crosstalk between epithelial and stromal cells in the differentiated organotypic state. Use of these models will aid in identification and validation of targets for development of novel therapeutics for treatment of human skin and airway epithelial diseases including cancers, chronic wounds, blistering diseases, scarring, and airway remodeling associated with asthma and COPD. Public Health Relevance: The gene modified tissue engineered models to be produced by the current project will provide researchers in the pharmaceutical industry and academic research laboratories a readily available means for studying the functional genomics of nearly any gene in human epithelial cells, as well as crosstalk between epithelial and stromal cells in the differentiated organotypic state. The models will be utilized to identify and validate targets for the development of novel therapeutics for treatment of human skin and airway epithelial diseases including cancers, chronic wounds, blistering diseases, scarring, and airway remodeling associated with asthma and COPD.